Question

Select the order in which the following steps should be performed to express a eukaryotic protein in bacterial cells.

Rank the options below.
1. Transform bacterial cells with the plasmid.
2. Identify a cDNA clone of the gene that encodes the protein.
3. Grow the bacterial cells to high density.
4. Add allolactose to the media.
5. Break open the bacterial cells and purify the eukaryotic protein.
6. Insert the cDNA downstream of the lac control region in a plasmid.

Answer 1

To express a eukaryotic protein in bacterial cells, the steps should be performed in the following order: identify the cDNA clone, insert the cDNA into a plasmid, transform bacterial cells, grow cells to high density, add allolactose, and then purify the protein.

Step-by-step explanation:

1. Identify a cDNA clone of the gene that encodes the protein.
2. Insert the cDNA downstream of the lac control region in a plasmid.
3. Transform bacterial cells with the plasmid.
4. Grow the bacterial cells to high density.
5. Add allolactose to the media.
6. Break open the bacterial cells and purify the eukaryotic protein.

First, the cDNA clone of the gene that encodes the protein needs to be identified. This can be done using techniques like reverse transcription and PCR. Then, the cDNA is inserted into a plasmid downstream of the lac control region to ensure expression. The plasmid is then transformed into bacterial cells, which are grown to high density. Allolactose is added to induce protein expression. Finally, the bacterial cells are broken open and the eukaryotic protein is purified.